Methods and compositions for promoting organ development

ABSTRACT

Compositions and methods are provided for promoting organ development in warm blooded animals, and in particular in certain aspects a premature infant or foetus. Compositions and methods are also provided for the administration of at least one colony stimulating factor-1 protein (CSF-1), precursor, variant, analogue, derivative thereof, or combinations thereof, or otherwise, at least one nucleic acid molecule encoding colony stimulating factor-1 protein (CSF-1), precursor, variant, analogue, derivative thereof, or combinations thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a continuation-in-part of a PCT applicationfiled in the Australian Receiving Office on Sep. 17, 2007, (which claimspriority to Australian Provisional Application No. AU2006905099, filedon 15 Sep. 2006, entitled “Method For Promoting Organ Development” andAustralian Provisional Application No. AU2006905156, filed 18 Sep. 2006,entitled “Method For Promoting Organ Development”), and is also acontinuation-in-part of International Patent Application No.PCT/AU2006/000357, filed Mar. 17, 2006, entitled “Renal Repair andRegeneration” (which claims priority to Australian ProvisionalApplication No. AU2005901346, filed 18 Mar. 2005, entitled “Renal Repairand Regeneration”). Each of these documents, including those inparenthesis, are incorporated herein by reference in their entirety. Inaddition, each of the following documents are incorporated herein byreference in its entirety:

Bertram J F (1995) Analyzing renal glomeruli with the new stereology;International Review of Cytology; 161:111-172.

Dressler, G. R. (2002). Development of the Excretory System.

Mouse Development—Patterning, Morphogenesis and Organogenesis.

J. Rossant and P. P. L. Tam. Houston, Academic Press:395; Goldenring J(2004). Respiratory Distress Syndrome in Infants. MedlinePlus MedicalEncyclopedia. [Available athttp://www.nlm.nih.gov/medlineplus/ency/article/001563.htm].

Hayashi M. Zhu K. Sagesaka T. Fukasawa I. Inaba N. Elevation of amnioticfluid macrophage colony-stimulating factor in normotensive pregnanciesthat delivered small-for-gestational-age infants. American Journal ofReproductive Immunology. 57(6):488-94, 2007 June.

Hinchliffe, S., Sargent, P., et al. (1991). “The effect of intrauterinegrowth expressed in absolute number of glomeruli assessed by the“disector” method and Cavalieri principle.” Lab Investigator 64:777-784.

Horster, M., Braun, G., et al. (1999). “Embryonic renal epithelia:Induction, nephrogenesis and cell differentiation.” PhysiologicalReviews 79(4):1157-1191.

Hume, D., Monkley, S., et al. (1995). “Detection of c-fms protooncogenein early mouse embryos by whole mount in situ hybridisation indicatesroles for macrophages in tissue remodelling.” British Journal ofHaematology 90(4):939-942.

Kett M M, Alcorn D, Bertram J F, Anderson W P (1996). Glomerulardimensions in spontaneously hypertensive rats: effects of AT1antagonism. Journal of Hypertension; 14:107-113.

Keith J C Jr. Pijnenborg R. Luyten C. Spitz B. Schaub R. Van Assche F A.Maternal serum levels of macrophage colony-stimulating factor areassociated with adverse pregnancy outcome. European Journal ofObstetrics, Gynecology, & Reproductive Biology. 89(1):19-25, 2000.

Wei S. Lightwood D. Ladyman H. Cross S. Neale H. Griffiths M. Adams R.Marshall D. Lawson A. McKnight A J. Stanley E R. Modulation ofCSF-1-regulated post-natal development with anti-CSF-1 antibody.Immunobiology. 210(2-4):109-19, 2005.

Dai X M. Zong X H. Sylvestre V. Stanley E R. Incomplete restoration ofcolony-stimulating factor 1 (CSF-1) function in CSF-1-deficientCsflop/Csflop mice by transgenic expression of cell surface CSF-1.Blood. 103(3):1114-23, 2004.

Seckl J R, Holmes M C. Mechanisms of disease: glucocorticoids, theirplacental metabolism and fetal ‘programming’ of adult pathophysiology.Nat Clin Pract Endocrinol Metab. 3(6):479-88, 2007.

Gennaro, Alfonso, Remington's Pharmaceutical Sciences, 18^(th) edition,Mack Publishing Co. (1990).

University of the Sciences in Philadelphia (editor) Remington: TheScience and Practice of Pharmacy 21^(st) edition (2005).

Rae F, Woods K, Sasmono T, Campanale N, Taylor D, Ovchinnikov D,Grimmond S M, Hume D A, Ricardo S D, and Little M H. Characterisationand trophic functions of murine embryonic macrophages based upon the useof a CSF-IR-EGFP transgenic reporter. Developmental Biology (In Press,accepted May 24) 2007.

FIELD OF THE INVENTION

The present disclosure relates to embodiments for promoting organdevelopment in warm blooded animals, and in particular in certainaspects a premature infant or foetus. Compositions and methods areprovided for the administration of colony stimulating factor-1 protein(CSF-1), or a precursor, variant, analogue or derivative thereof, orotherwise, a nucleic acid molecule encoding colony stimulating factor-1protein (CSF-1), or a precursor, variant, analogue or derivativethereof.

BACKGROUND OF THE INVENTION

Compared to infants who have born following a normal, full termpregnancy, premature infants, particularly babies born before 32 weeksof gestation, are at a considerably greater risk of developing a numberof serious health problems including, for example, renal and lungdisorders.

For instance, the low birth weight and insufficient physical developmentof premature infants predisposes them to respiratory complications suchas respiratory distress syndrome (RDS) and chronic lung disease (alsoknown as bronchopulmonary dysplasia). RDS is associated with irregularbreathing difficulties and occurs in approximately 60 to 80 percent ofinfants born before 28 weeks gestation, and in 15 to 30 percent of thoseborn between 32 and 36 weeks of gestation. Treatment of such infantstypically involves supplemental oxygen, but in some cases, also requiresthe use of a mechanical ventilator and continuous positive airwaypressure. Moreover, in severe cases, treatment will additionally involvethe administration of an artificial lung surfactant. While suchtreatments are very successful, long-term ventilator treatment isundesirable since this can lead to lung deterioration, which in turn,can lead to bronchopulmonary dysplasia.

It is also known that premature infants are born with reduced numbers ofnephrons (filtration units of the kidney), an outcome that may beassociated with increased risk of developing hypertension and reducedrenal function following injury later in life.

Lung Development: Analogies Between Human and Mouse

The human lung is derived from the foregut at about 4 weeks gestationand begins as a diverticulum. The lung diverticulum is covered withsplanchnic mesoderm that gives rise to the tissue components of themature adult lung such as cartilage, smooth muscle and blood vessels.Lung development is characterised by branching morphogenesis that givesrise to the primary, secondary and tertiary bronchi. The stages offoetal lung development are classified into three distinct phases,namely; the pseudoglandular, canalicular and saccular phases. Someaspects of alveolar lung development including epithelial celldifferentiation begin in the canalicular phase. However, approximately15-18% of alveoli form late in gestation, with most of the alveoliformed after birth. Shortly after birth, the surface area of theair-blood interface increases with the formation of the alveolar ductsand sacs.

Premature infants can survive with lung development in the latecanalicular or early saccular phase. This is a phase when the conductingairways have stopped branching and are enlarging at their distaltermini. There is a progressive loss of extracellular matrix andmesenchymal cells that separate the capillaries from the sites ofalveoli. These premature infants survive without alveoli by treatmentinvolving mechanical ventililation and the administration of anartificial lung surfactant, although, as mentioned above, they are atrisk of developing bronchopulmonary dysplasia.

In mice, the lung also arises from the ventral foregut, but atapproximately embryonic day 9.5 (E9.5). Subsequently, the respiratorytree develops through the pseudoglandular (E9.5-16.5), canalicular(E16.5-17.5), and saccular (E17.5-postnatal day 5) phase. While mouseand human lung development is highly analogous from an embryologicalpoint of view and while the same genes are critical in both organisms,in contrast to the human lung, alveolarisation is not complete beforebirth in the mouse. At birth, the mouse lungs consists of immatureterminal saccules with some secondary septa, with alveolarisation andalveolar separation occurring during the during the first 1-3 postnatalweeks. The alveolar surfaces increase through the enlargement ofpre-existing alveoli with formation of new alveoli.

Kidney Development: Analogies Between Humans and Mice

The development of the kidney is highly analogous between human andmouse with respect to the embryological origin of the tissues involved,the morphogenetic processes and the genes regulating these processes.

In the human (as for the mouse), both the renal and genital systemsoriginate from the intermediate mesoderm. Development of the kidneyundergoes three distinct stages before resulting in the mature adultkidney. The process begins with the formation of the pronephros, thenthe mesonephros and finally the metanephros, after which the pronephrosand mesonephros regress, and the metanephros remains to form thefunctional adult kidney. Metanephric development begins with theoutgrowth of ureteric bud, originating from the Wollfian duct, invadingthe surrounding metanephric mesenchyme. The functional units within thekidney responsible for filtration of the blood, concentration of thefiltrate to form urine and reclamation of water and ions are thenephrons. The formation of these functional units is referred to asnephrogenesis. Human nephrogenesis (development of kidney nephrons) iscompleted before birth. The number of nephrons in normal human kidneysranges from approximately 300,000 to more than one million. After birth,the nephron number is complete and no new nephrons are able to beformed. In humans, development of the permanent kidney begins aroundgestational week 5. In the third trimester, 60% of nephrons are formedand continue to form until approximately 36 weeks. No new nephrons areformed after this time.

In the mouse (as with humans), there are three embryonic kidneys, thepronephros, mesonephros and metanephros, and the development of thefinal permanent kidney, the metanephros, begins with the outgrowth ofureteric bud, originating from the Wollfian duct, invading thesurrounding metanephric mesenchyme. This occurs at around embryonic day9-10.5 (E9-10.5) and requires inductive signals from the metanephricmesenchyme to initiate bud development. The induced mesenchyme sendsreciprocal signals to induce growth and branching of the ureteric bud.Nephron formation (nephrogenesis) is induced when factors secreted bythe ureteric bud cause the induction, condensation and aggregation ofthe mesenchyme. Each aggregate undergoes epithelialisation and thenproceeds through the developmental stages of the polarised vesiclestage, the comma and the S-stage. There is continued branching with newaggregates forming at the tips, and this process continues with theinduction of new nephrons. By the end of nephrogenesis, there are morethan 26 terminally differentiated cell types with distinct location,morphology and function. Unlike the human, in the mouse kidneydevelopment continues in mice until around 7-10 days after birth.

Growth Factors in Kidney and Lung Development

Growth factors, aside from their influence in cell growth, contributegreatly to many processes including cell migration, morphogenesis,differentiation and proliferation. The roles of growth factors inbranching morphogenesis in the lung and nephrogenesis in the kidney arecontrolled by an array of inductive and inhibitory signals. The crucialroles of factors including insulin-like growth factor-I and II (IGF-Iand IGF-II), hepatocyte growth factor (HGF), and epithelial growthfactor (EGF) have been well established in the developing lung andkidney. It is, however, considered that there may be numerous othergrowth factors which play significant roles in development of the lungand kidney.

In has been found that in warm blooded animals, usings the embodimentsdisclosed it is possible to promote organ development (as reflected in,for some organs, an increase in organ weight), and more particularly,increased growth and/or enhanced nephrogenesis and lung maturation. Ithas also been found that promoting organ development and/or maturationin a warm blooded premature infant or foetus is possible.

SUMMARY OF THE INVENTION

Certain embodiments disclosed provide compositions for and methods fortreating complications arising from or related to low birth weight inmammals, including for example, humans, pigs, horses, dogs and otherlivestock. Low birth weight may be caused by premature or preterm birthor by poor foetal growth, such as intrauterine growth restriction. Thereare many causes of poor foetal growth, some of which include chromosomalabnormalities, placental dysfunction, placenta previa, smoking, drug oralcohol abuse, amnionitis, abruptio placentae or preeclampsia, maternalhypertension, maternal hypoxemia, maternal toxemia, polyhydramnios,urinary tract infection, malnutrion, infection, anemia, diabetes,inadequate maternal weight gain and various diseases. The compositionsand methods described herein may be used to treat and/or prevent any ofthese causes of low birth weight and the complications. In someembodiments, the compositions and methods described herein may be usedspecifically to treat or prevent causes of low birth weight such asfoetal alcohol syndrome, placental insufficiency, intrauterine growthretardation (IUGD), foetal growth restriction as a result of infections,genetic abnormalities such as mutations in the gene that encodes11-β-hydroxysteroid dehydrogenase type 2, maternal hypertension,diabetes, alcohol and illicit drug abuse or inadequate maternal weightgain.

Low birth weight in babies can result in a large number of complicationsincluding immature organ growth, such as immature lungs and kidneys,respiratory distress syndrome (RDS), intraventricular hemmorhage (IVH),Patent ductus arteriosus (PDA), necrotizing enterocolitis (NEC),retinopathy of prematurity (ROP), and osteopenia. The long term adverseeffects of a low birth weight include increased risk of heart diseaseand renal failure, increased risk of diabetes and obesity and a possibleconsequence on intelligence. The compositions and methods describedherein may be used to treat one or more complications arising from lowbirth weight in mammals and may be administered prior to birth, such asto the mother or to the foetus or after birth, such as to the infant.

Certain embodiments disclosed provide methods of treating complicationsarising from or related to low birth weight in mammals, such as inhumans, pigs, dogs, horses or other livestock, such as in prematureinfants, in low birth weight infants or in foetuses comprisingadministering to said mammals:

at least one colony stimulating factor-1 protein (CSF-1), and/or atleast one precursor, variant, analogue , derivative thereof orcombination thereof, or

at least one nucleic acid molecule encoding said at least one colonystimulating factor-1 protein (CSF-1), and/or at least one precursor,variant, analogue, derivative thereof, or combination thereof.

Certain embodiments disclosed include methods of treating complicationsarising from or related to low birth weight in mammals such as inhumans, pigs, horses, dogs or other livestock, such as in prematureinfants, in low birth weight infants or in foetuses comprisingadministering to said mammals:

a low birth weight complications-reducing or-limiting amount of at leastone colony stimulating factor-1 protein (CSF-1), and/or at least oneprecursor, variant, analogue, derivative thereof, or combinationthereof, or

a therapeutically effective amount of at least one nucleic acid moleculeencoding a low birth weight complications-reducing or-limiting amount ofsaid at least one colony stimulating factor-1 protein (CSF-1), and/or atleast one precursor, variant, analogue, derivative thereof orcombination thereof.

Certain embodiments disclosed provide pharmaceutical compositions fortreating, reducing or limiting complications arising from or related tolow birth weight in mammals, such as in humans, pigs, horses, dogs orother livestock, such as in premature infants, in low birth weightinfants or in foetuses comprising administering to said mammals:

a low birth weight complications-reducing or-limiting amount of at leastone colony stimulating factor-1 protein (CSF-1), and/or at least oneprecursor, variant, analogue, derivative thereof or combination thereof,or

a therapeutically effective amount of at least one nucleic acid moleculeencoding a low birth weight complications-reducing or-limiting amount ofsaid at least one colony stimulating factor-1 protein (CSF-1), and/or atleast one precursor, variant, analogue, derivative thereof orcombination thereof.

Certain embodiments disclose methods of promoting organ developmentand/or maturation in mammals, such as in humans, pigs, dogs, horses orother livestock, such as in premature infants, in low birth weightinfants or in foetuses are provided. In certain aspects, methods ofpromoting organ development and/or maturation in mammals are disclosedthat comprise the step of administering to the mammal such as the human,pig, horse or other livestock, such as the premature infant, low birthweight infant or foetus:

colony stimulating factor-1 protein (CSF-1), and/or a precursor,variant, analogue, derivative thereof, or combination thereof or

a nucleic acid molecule encoding said colony stimulating factor-1protein (CSF-1), and/or a precursor, variant, analogue, derivativethereof or combination thereof.

In certain aspects, methods of promoting organ development and/ormaturation in mammals are disclosed that comprise the step ofadministering to the mammal such as the human, pig, horse, dogs or otherlivestock, such as the premature infant, low birth weight infant orfoetus:

at least one colony stimulating factor-1 protein (CSF-1), and/or atleast one precursor, variant, analogue, derivative thereof orcombination thereof, or

at least one nucleic acid molecule encoding said colony stimulatingfactor-1 protein (CSF-1), and/or at least one a precursor, variant,analogue, derivative thereof or combination thereof.

In certain aspects, the methods of promoting organ development in apremature infant, in a low birth weight infant or in a foetus disclosedcomprise the step of administering to the premature infant, the lowbirth weight infants or the foetus:

a premature infant, a low birth weight infant or a foetus organdevelopment-enhancing amount of at least one colony stimulating factor-1protein (CSF-1), and/or at least one precursor, variant, analogue,derivative thereof or combination thereof, or

a therapeutically effective amount of at least one nucleic acid moleculeencoding a premature infant, a low birth weight infant or a foetus organdevelopment-enhancing amount of said at least one colony stimulatingfactor-1 protein (CSF-1), and/or at least one precursor, variant,analogue, derivative thereof or combination thereof.

Certain embodiments disclosed provide pharmaceutical compositions forpromoting organ development in a premature infant, in a low birth weightinfant or in a foetus that comprise

a premature infant, a low birth weight infant or a foetus organdevelopment-enhancing amount of at least one colony stimulating factor-1protein (CSF-1), and/or at least one precursor, variant, analogue,derivative thereof or combination thereof, or

a therapeutically effective amount of at least one nucleic acid moleculeencoding a premature infant, a low birth weight infant or a foetus organdevelopment-enhancing amount of said at least one colony stimulatingfactor-1 protein (CSF-1), and/or at least one precursor, variant,analogue, derivative thereof or combination thereof.

In certain aspects, methods are disclosed that promote growth and/orenhance lung development and/or maturation in mammals, such as inhumans, pigs, horses, dogs or other livestock, such as in prematureinfants, in low birth weight infants or in foetuses. In certain aspects,the methods of promoting lung growth and/or enhancing lung developmentand/or maturation in mammals, such as in humans, pigs, horses, dogs orother livestock, such as in premature infants, in low birth weightinfants or in foetuses comprising administering to said mammals::

colony stimulating factor-1 protein (CSF-1), and/or a precursor,variant, analogue, derivative thereof or combination thereof, or

a nucleic acid molecule encoding said colony stimulating factor-1protein (CSF-1), and/or a precursor, variant, analogue, derivativethereof or combination thereof.

In certain aspects, the methods of promoting lung growth and/orenhancing lung development and/or maturation in a premature infant, in alow birth weight infant or in a foetus disclosed comprise the step ofadministering to the infant or foetus:

at least one colony stimulating factor-1 protein (CSF-1), and/or atleast one precursor, variant, analogue, derivative thereof, orcombination thereof, or

at least one nucleic acid molecule encoding said at least one colonystimulating factor-1 protein (CSF-1), and/or at least one precursor,variant, analogue, derivative thereof or combination thereof.

In certain aspects, the methods of promoting lung growth and/orenhancing lung development and/or maturation in a premature infant, in alow birth weight infant or in a foetus disclosed comprise the step ofadministering to the infant or foetus:

a premature infant, a low birth weight infant or a foetus lung growthpromoting and/or lung development and/or maturation-enhancing amount ofat least one colony stimulating factor-1 protein (CSF-1), and/or atleast one precursor, variant, analogue, derivative thereof orcombination thereof, or

a therapeutically effective amount of at least one nucleic acid moleculeencoding a premature infant, a low birth weight infant or a foetus lunggrowth promoting and/or lung development and/or maturation-enhancingamount of said at least one colony stimulating factor-1 protein (CSF-1),and/or at least one precursor, variant, analogue, derivative thereof orcombination thereof.

Certain embodiments disclosed provide pharmaceutical compositions forpromoting lung growth and/or enhancing lung development and/ormaturation in a premature infant, in a low birth weight infant or in afoetus that comprise

a premature infant, a low birth weight infant or a foetus lung growthpromoting and/or lung development and/or maturation-enhancing amount ofat least one colony stimulating factor-1 protein (CSF-1), and/or atleast one precursor, variant, analogue, derivative thereof orcombination thereof, or

a therapeutically effective amount of at least one nucleic acid moleculeencoding a premature infant, a low birth weight infant or a foetus lunggrowth promoting and/or lung development and/or maturation-enhancingamount of said at least one colony stimulating factor-1 protein (CSF-1),and/or at least one precursor, variant, analogue, derivative thereof orcombination thereof.

In certain embodiments, methods of promoting growth, maturation and/orenhancing kidney development in mammals, such as in humans, pigs,horses, dogs or other livestock, such as in premature infants, in lowbirth weight infants or in foetuses. In certain aspects the methods ofpromoting growth, maturation and/or enhancing kidney development inmammals, such as in humans, pigs, horses, dogs or other livestock, suchas in premature infants, in low birth weight infants or in foetusescomprising administering to said mammals:

colony stimulating factor-1 protein (CSF-1), and/or a precursor,variant, analogue, derivative thereof or combination thereof, or

a nucleic acid molecule encoding said colony stimulating factor-1protein (CSF-1), and/or a precursor, variant, analogue, derivativethereof or combination thereof.

In certain aspects the methods of promoting growth, maturation and/orenhancing kidney development in a premature infant, in a low birthweight infant or in a foetus comprise the step of administering to theinfant or foetus;

at least one colony stimulating factor-1 protein (CSF-1), and/or atleast one precursor, variant, analogue, derivative thereof, orcombination thereof, or

at least one nucleic acid molecule encoding said colony stimulatingfactor-1 protein (CSF-1), and/or at least one precursor, variant,analogue, derivative thereof or combination thereof.

In certain aspects, the methods of growth, maturation and/or enhancingkidney development in a premature infant, in a low birth weight infantor in a foetus disclosed comprise the step of administering to theinfant or foetus:

a premature infant, a low birth weight infant or a foetus kidney growthmaturation and/or development-enhancing amount of at least one colonystimulating factor-1 protein (CSF-1), and/or a precursor, variant,analogue, derivative thereof or combination thereof, or

a therapeutically effective amount of at least one nucleic acid moleculeencoding a premature infant, a low birth weight infant or a foetuskidney growth maturation and/or development-enhancing amount of said atleast one colony stimulating factor-1 protein (CSF-1), and/or at leastone precursor, variant, analogue, derivative thereof or combinationthereof.

Certain embodiments disclosed provide pharmaceutical compositions forpromoting growth, maturation and/or enhancing kidney development in apremature infant, in a low birth weight infant or in a foetus thatcomprise

a premature infant, a low birth weight infant or a foetus kidney growthmaturation and/or development-enhancing amount of at least one colonystimulating factor-1 protein (CSF-1), and/or at least one precursor,variant, analogue, derivative thereof or combination thereof, or

a therapeutically effective amount of at least one nucleic acid moleculeencoding a premature infant, a low birth weight infant or a foetuskidney growth maturation and/or development-enhancing amount of at leastone said colony stimulating factor-1 protein (CSF-1), and/or at leastone precursor, variant, analogue, derivative thereof or combinationthereof.

In certain aspects, methods are disclosed that promote growth and/orenhance bone development and/or maturation in mammals, such as inhumans, pigs, horses, dogs or other livestock, such as in prematureinfants, in low birth weight infants or in foetuses. In certain aspects,the methods of promoting bone growth and/or enhancing bone developmentand/or maturation in mammals, such as in humans, pigs, horses, dogs orother livestock, such as in premature infants, in low birth weightinfants or in foetuses comprising administering to said mammals:

colony stimulating factor-1 protein (CSF-1), and/or a precursor,variant, analogue, derivative thereof or combination thereof or

a nucleic acid molecule encoding said colony stimulating factor-1protein (CSF-1), and/or a precursor, variant, analogue, derivativethereof or combination.

In certain aspects, the methods of promoting bone growth and/orenhancing bone development and/or maturation in a premature infant, in alow birth weight infant or in a foetus disclosed comprise the step ofadministering to the infant or foetus:

at least one colony stimulating factor-1 protein (CSF-1), and/or atleast one precursor, variant, analogue, derivative thereof orcombinations thereof or

at least one nucleic acid molecule encoding said at least one colonystimulating factor-1 protein (CSF-1), at least one precursor, at leastone variant, at least one analogue, at least one derivative thereof orcombinations thereof.

In certain aspects, the methods of promoting bone growth and/orenhancing bone development and/or maturation in a premature infant, in alow birth weight infant or in a foetus disclosed comprise the step ofadministering to the infant or foetus:

a premature infant, a low birth weight infant or a foetus bone growthpromoting and/or bone development and/or maturation-enhancing amount ofat least one colony stimulating factor-1 protein (CSF-1), and/or atleast one precursor, variant, analogue, derivative thereof orcombination thereof or

a therapeutically effective amount of at least one nucleic acid moleculeencoding a premature infant, a low birth weight infant or a foetus bonegrowth promoting and/or bone development and/or maturation-enhancingamount of said at least one colony stimulating factor-1 protein (CSF-1),and/or at least one precursor, variant, analogue, derivative thereof orcombination thereof.

Certain embodiments disclosed provide pharmaceutical compositions forpromoting bone growth and/or enhancing bone development and/ormaturation in a premature infant, in a low birth weight infant or in afoetus that comprise

a premature infant, a low birth weight infant or a foetus bone growthpromoting and/or bone development and/or maturation-enhancing amount ofat least one colony stimulating factor-1 protein (CSF-1), and/or aprecursor, variant, analogue, derivative thereof or combination thereof,or

a therapeutically effective amount of at least one nucleic acid moleculeencoding a premature infant, a low birth weight infant or a foetus bonegrowth promoting and/or bone development and/or maturation-enhancingamount of said at least one colony stimulating factor-1 protein (CSF-1),and/or at least one precursor, variant, analogue, derivative thereof orcombination thereof.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 provides a graph demonstrating the average mouse body weight(n=3/group) in mice receiving CSF-1 compared to litter mate controltreated mice.

FIG. 2 provides a graph showing the effect on kidney weight of CSF-1delivery to newborn mouse pups.

FIG. 3 provides a graph showing a stereological estimation of glomerularnumber in the kidneys from mice receiving CSF-1 or phosphate bufferedsaline (PBS).

FIG. 4 shows the histology of kidneys from control (A; Mag ×100) andCSF-1 treated mice (B; Mag ×100) killed at day 29 and stained withhaematoxylin and eosin; and the histology of lungs of control (C; Mag×200, E; Mag ×400) and CSF-1-treated (D; Mag ×200, F; Mag ×400) mice.

DETAILED DESCRIPTION

In general, because of the strong similarities between all placentalmammals in terms of organogenesis, the mouse provides an excellentpredictive model for organogenesis in humans, pigs, horses, dogs andother placental mammals.

Growth factor known as colony stimulating factor-1 protein (CSF-1)(alsoknown as macrophage colony stimulating factor (M-CSF)) controls thesurvival, proliferation and differentiation of cells of themonocyte/macrophage lineage, and acts by binding to the CSF-1 receptor(CSF-1R), a cell-surface tyrosine kinase receptor encoded by the c-fmsproto-oncogene. Previous studies have shown that c-fms mRNA is found inthe placenta, localised to cells of a macrophage specific lineage (Hume,Monkley et al., 1995). The present embodiments relate to and elucidatethe role(s) that CSF-1 has in embryonic development. The applicants havefound, surprisingly, that in newborn mice, CSF-1 was able to treatcomplications arising from or related to low birth weight, to promoteorgan development, and more particularly, increased growth and/orenhanced lung maturation and nephrogenesis in the kidney and increasedbone growth and/or enhanced bone maturation in co-occurrence with anoverall increase in size and body weight. Lung and kidney development isincomplete in newborn mice, newborn mice, therefore, provide a usefulmodel for lung and kidney development in the human foetus and prematureinfants. In addition, newborn mice undergo bone and cartilageremodelling and growth postnatally. Thus, as disclosed herein theadministration of CSF-1 to premature infants and pregnant mothers atrisk of premature birth (or for whom premature birth is desirable) maypermit treatment and/or prevention of diseases and conditions associatedwith underdeveloped organs such as the lungs and kidneys and boneformation.

In certain embodiments, based on studies conducted to elucidate whatrole(s) CSF-1 might have in embryonic development, the presentapplicants surprisingly found that in certain embodiments in newbornmice, CSF-1 was able to promote organ development (as reflected in, forsome organs, an increase in organ weight), and more particularly,increased growth and/or enhanced nephrogenesis and lung maturation.

In certain embodiments, methods of promoting organ development and/ormaturation in a premature infant or foetus, the methods comprising thestep of administering to the infant or foetus;

colony stimulating factor-1 protein (CSF-1), or a precursor, variant,analogue or derivative thereof, or

a nucleic acid molecule encoding said colony stimulating factor-1protein (CSF-1), or a precursor, variant, analogue or derivativethereof.

The methods disclosed herein may be used to treat complications arisingfrom or related to low birth weight, to promote the development of oneor more organs such as, but not limited to, the lung, kidney, brain, eyeand organs of the gastrointestinal (G.I.) tract, in certain aspects inparticular the small intestine, and may be used to promote bone growthand development in mammals, such as humans, pigs, horses, dogs and otherlivestock, such as in premature infants or in foetuses. The organdevelopment that may be achieved by the method disclosed herein canresult in cell growth and cell differentiation so as to cause organmaturation (e.g. in terms of organ structure and function) towards thatof infants born following a normal, full-term pregnancy and foetaldevelopment. The premature infant or foetus treated in accordance withcertain embodiments may thereby avoid or defer, for example, developinghypertension and/or reduced renal function following injury later inlife, respiratory distress syndrome (RDS) and bronchopulmonarydysplasia, intraventricular hemmorhage and neural development disordersthat can lead to learning problems, behavioural problems and cerebralpalsy, retinopathy due to abnormal growth of blood vessels, and hearingloss.

In certain embodiments, the methods involve the administration of CSF-1,at least one nucleic acid encoding CSF-1, or combinations thereof. Incertain preferred embodiments, the methods disclosed involve theadministration of human CSF-1, a least one nucleic acid encoding humanCSF-1, or combinations thereof. However, it is also suitable in certainembodiments to administer a precursor, variant, analogue or derivativeof CSF-1, at least one nucleic acid encoding same, or combinationsthereof. In certain aspects it is preferred to administer, at least oneprecursor, at least one variant, at least one analogue, at least onederivative of human CSF-1, at least one nucleic acid encoding same, orcombinations thereof.

The term “precursor” is to be understood to refer to any molecule thatis converted or metabolised within the body to CSF-1. Thus, one exampleof a suitable CSF-1 precursor is an immature CSF-1 comprising itsnative, or a heterologous, secretory signal, which can be processed byproteolytic cleavage to produce CSF-1 (i.e. mature CSF-1).

The term “variant” is to be understood to refer to an isoform of CSF-1encoded by, for example, an allelic variant.

The term “analogue” is to be understood to refer to any molecule thatdiffers from CSF-1 but retains similarity, or substantial similarity, inbiological function of CSF-1, in particular the ability to promote organdevelopment. In certain aspects, an analogue may have substantialoverall structural similarity with CSF-1 or only structural similaritywith one or more regions or domains of CSF-1 responsible for itsbiological function. Typically, an analogue of CSF-1 will be providedby, or be the result of, the addition of one or more amino acids to theamino acid sequence of CSF-1, deletion of one or more amino acids fromthe amino acid sequence of CSF-1, and/or substitution of one or moreamino acids of the amino acid sequence of CSF-1, and/or combinationsthereof. In certain aspects, inversion of amino acids and othermutational changes that result in the alteration of the amino acidsequence are also encompassed. Such an analogue may be prepared byintroducing nucleotide changes into a nucleic acid molecule such thatthe desired amino acid changes are achieved upon expression of themutagenised nucleic acid molecule, or by otherwise synthesising an aminoacid sequence incorporating the desired amino acid changes. Thesubstitution of an amino acid may involve conservative ornon-conservative amino acid substitution. By conservative amino acidsubstitution, it is meant that an amino acid residue is replaced withanother amino acid having similar, or substantially similar,characteristics and which does not substantially alter the desiredbiological function of the protein. Exemplary conservative amino acidsubstitutions are provided in Table 1 below. In certain aspects,particular conservative substitutions envisaged are: G, A, V, I, L, M;D, E, N, Q; S, C, T; K, R, H; and P, N-α-alkylamino acids. In certainaspects, conservative amino acid substitutions may be selected on thebasis that they do not have any substantial effect on (a) the structureof the peptide backbone in the region of the substitution, (b) thecharge or hydrophobicity of the protein at the site of substitution, (c)the bulk of the side chain at the site of substitution, and/orcombinations thereof.

TABLE 1 Exemplary conservative amino acid substitutions ConservativeSubstitutions Ala Val*, Leu, Ile Arg Lys*, Gln, Asn Asn Gln*, His, Lys,Arg, Asp Asp Glu*, Asn Cys Ser Gln Asn*, His, Lys, Glu Asp*,γ-carboxyglutamic acid (Gla) Gly Pro His Asn, Gln, Lys, Arg* Ile Leu*,Val, Met, Ala, Phe, norleucine (Nle) Leu Nle, Ile*, Val, Met, Ala, PheLys Arg*, Gln, Asn, ornithine (Orn) Met Leu*, Ile, Phe, Nle Phe Leu*,Val, Ile, Ala Pro Gly*, hydroxyproline (Hyp), Ser, Thr Ser Thr Thr SerTrp Tyr Tyr Trp, Phe*, Thr, Ser Val Ile, Leu*, Met, Phe, Ala, Nle*indicates preferred conservative substitutions

In certain aspects, where an analogue is prepared by synthesis, theanalogue may also include an amino acid or amino acids not encoded bythe genetic code, such as γ-carboxyglutamic acid and hydroxyproline. Forexample, D-amino acids rather than L-amino acids may be included. A listof amino acids not encoded by the genetic code is provided in Table 2.In a certain preferred embodiments, the analogue is a mimetic of CSF-1such as a peptido-mimetic. However, it is not always necessary that ananalogue of CSF-1 have amino acid sequence identity and/or similarity.In certain aspects an analogue may not be proteinaceous at all. Incertain embodiments an analogue may have at least 75%, such as at least80%, at least 85%, at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98% or at least 99% homology with CSF-1.

TABLE 2 List of amino acids not encoded by the genetic codeα-aminobutyric acid D-α-methylhistidine L-N-methyl-t- butylglycineα-amino-α- D-α-methylisoleucine L-norleucine methylbutyrateAminocyclopropane- D-α-methylleucine L-norvaline carboxylateAminoisobutyric acid D-α-methyllysine α-methyl- aminoisobutyrateAminonorbornyl- D-α- α-methyl-α- carboxylate methylmethionineaminobutyrate Cyclohexylalanine D-α- α-methylcyclohexyl methylornithinealanine Cyclopentylalanine D-α- α-methylcylcopentyl methylphenylalaninealanine L-N-methylisoleucine D-α-methylproline α-methyl-α-napthylalanine D-alanine D-α-methylserine α-methylpenicillamine D-arginine D-α-N-(4-aminobutyl)glycine methylthreonine D-aspartic acid D-α-N-(2-aminoethyl)glycine methyltryptophan D-cysteine L-N-methylalanineN-(3- aminopropyl)glycine D-glutamate L-N-methylarginineN-amino-α-methyl butyrate D-glutamic acid L-N- α-napthylalaninemethylasparagine D-histidine L-N-methylaspartic N-benzylglycine acidD-isoleucine L-N-methylcysteine N-(2-carbamylediyl) glycine D-leucineL-N- N-(carbamylmethyl) methylglutamine glycine D-lysineL-N-methylglutamic N-(2- acid carboxyethyl)glycine D-methionineL-N-methylhistidine N- (carboxymethyl)glycine D-ornithineL-N-methylleucine N-cyclobutylglycine D-phenylalanine L-N-methyllysineN-(N-(3,3- diphenylpropyl carbamylmethyl)glycine D-proline L-N-N-(N-(2,2-diphenylethyl methylmethionine carbamylmethyl)glycine D-serineL-N- 1-carboxy-1-(2,2- methylnorleucine diphenyl-ethylamino)cyclopropane D-threonine L-N-methylnorvalineL-α-methyltryptophan D-tryptophan L-N-methylornithineN-cycloheptylglycine D-tyrosine L-N- N-cyclohexylglycinemethylphenylalanine D-valine L-N-methylproline N-cyclodecylglycineD-α-methylalanine L-N-medlylserine L-α-methylnorleucineD-α-methylarginine L-N-methylthreonine L-α-methylornithine D-α- L-N-L-α-methylproline methylasparagine methyltryptophan D-α-methylaspartateL-N-methyltyrosine L-α-methylthreonine D-α-methylcysteineL-N-methylvaline L α-methyltyrosine D-α-methylglutamine L-N-L-N-methylhomo- methylethylglycine phenylalanine D-α-methyltyrosineL-α-methylleucine L-α-methylserine L-α- L-α-methyllysine L-α-methylmethionine methylphenylalanine L-α-methylnorvatineL-α-methylvaline

The term “derivative” is to be understood to refer to any molecule thatis derived (substantially derived) or obtained (substantially obtained)from CSF-1, but retains similarity, or substantial similarity, inbiological function of CSF-1. In certain aspects, the biologicalfunction is the ability to promote organ development. A derivative may,for instance, be provided as a result of cleavage of CSF-1 to producebiologically-active fragments, cyclisation, bioconjugation and/orcoupling with one or more additional moieties that improve, for example,solubility, stability or biological half-life, or which act as a labelfor subsequent detection or the like. A derivative may also result frompost-translational or post-synthesis modification such as the attachmentof carbohydrate moieties, or chemical reaction(s) resulting instructural modification(s) such as alkylation or acetylation of an aminoacid(s) or other changes involving the formation of chemical bonds. In aparticularly preferred embodiment of a derivative suitable for use inthe present invention, the derivative is the mature domain of CSF-1. Inanother preferred embodiment of a derivative suitable for use in themethods disclosed herein, the derivative is a biologically active,C-terminal fragment of CSF-1 (e.g. a CSF-1 fragment comprising theC-terminal amino acids 1 to 150 of the 536 amino acid protein). Furtherembodiments of a derivative of CSF-1 include CSF-1 comprising chemicallymodified side chains (e.g. pegylation of lysyl ε-amino groups), C-and/or N-termini (e.g. acylation of the N-terminal with aceticanhydride), or linked to various carriers (e.g. human serum albumin orhistidine (His₆) tag).

CSF-1 produced from synthetic protein synthesis and chemical ligationmay be used as a source for delivery of large amounts of protein toanimals or infants. These synthesized protein analogues may haveimproved potency or pharmacokinetic properties in comparison to naturalCSF-1. CSF-1 protein may be made by first making individual peptidesegments of the protein using solid-phase peptide synthesis (SPPS) andthen after purification, joining the segments chemically, or vialigatation, in solution to form the full-length polypeptide. Tofacilitate the ligation of individual peptide segments an N-terminalcysteine residue (generally occurring naturally in the protein sequence)and a C-terminal thioester (prepared on-resin) may be needed. Aftersynthesis, the unfolded full-length CSF-1 polypeptide may be folded intoits biologically active conformation.

Preferably, certain embodiments disclosed involve the administration ofat least one recombinant human CSF-1 (rhCSF-1), in particular,bacterially-expressed, non-glycosylated recombinant human CSF-1.

CSF-1, or a precursor, variant, analogue or derivative thereof, may beadministered to the premature infant or foetus by any effective method,some of which are known. For example, for the premature infant, theroute of administration can be selected from, for example, intramuscular(i.m.), intravenous (i.v.), topical, such as inhalationaladministration, intratracheal, subcutaneous (s.c.) administration and/orcombinations thereof. On the other hand, for the foetus, the route ofadministration may be selected from i.m., i.v., s.c., intrauterine(i.u.), oral, inhalational administration to the pregnant mother, and/orcombinations thereof.

In some embodiments, rather then systemic administration, it may bedesirable for the route of administration to be localized to one or morespecific organs or portions of the body, such as by direct applicationof the therapeutic to the target treatment area or areas. In someembodiments, the composition may be an immediate release dosage form. Inother embodiments the composition may be a time release dosage form,including an implantable controlled release form. In some embodiments,the dosage forms may include tablets, dispersions, suspensions,solutions, injections, syrups, troches, capsules suppositories,aerosols, transdermal patches and the like.

Certain embodiments may be administered to ventilated premature infantsusing aerosol delivery. “Preterm” or “premature” birth can be defined asdelivery before approximately the thirty-seventh week of pregnancy.Preterm deliveries can be further delineated as either “very preterm”(before approximately the thirty-third week) or “moderately preterm”(between the approximately thirty-third and approximately thethirty-sixth weeks). Mechanical ventilation that is heated ornon-heated; and humidified or non-humidified may be performed ininfants. Because of the small tidal volumes and high respiratory ratesrequired for an infant, ventilation may be time or pressure cycled, witha continuous flow of gas circulating through the ventilator circuit.

Certain embodiments of the composition and methods disclosed may beinclude delivery of CSF-1 with ventilation by aerosol delivery usingeither a vibrating mesh nebulizer, a jet nebuliser, a metered doseinhaler (MDI), an ultrasonic nebulizer, or an electric pump nebuliser.For example, CSF-1 may be administered in the nebulizer as a bolus dosebefore the initiation of positive pressure ventilation. In anotherexample CSF-1 may be delivered using continuous feed through an infusionset into a nebulizer. Such a method will allow dosing of aerosol atdifferent rates by adjusting the flow of the drug/unit of time into thenebulizer. Nebulization of certain embodiments disclosed herein mayensure more effective drug delivery to, for example, the lung alveoli ofthe premature infants.

In certain embodiments, nebulization treatment may be delivered as soonas possible after birth and delivered either intermittently or withcontinuous aerosol therapy. Continuous nebulization therapy may involvethe delivery of prescribed dose of, for example, CSF-1 in diluents orsterile saline or phosphate buffered-saline over 8 hour periods. CSF-1may be delivered with a small volume-limited or large-volume nebuliserwith infusion pump. The volume of CSF-1 to be delivered to infants inthe nebulizer may be in the range of 5-15 ml. The CSF-1 may be added tothe pediatric nebulizer unit in the inspiratory limb of the ventilatorcircuit about 10-30 cm away from the patient wye.

The nebulisers may be placed in the ventilator manifold and set todeliver a CSF-1 to an infant at a dose ranging from 0.01-1000 μg/hour ofCSF-1 continuously over 8 hours for 1 day, 2 days, 3 days, 4 days, 5days, 6 days or 7 consecutive days. Typically, such an amount may be, inthe case of administration to the premature infant, in the range ofabout 0.1 to 500 μg/h, about 0.05 to 500 μg/h, about 0.2 to 400 μg/h,0.1 to 1000 μg/h, about 0.05 to 1000 μg/h, about 0.1 to 1000 μg/h, about0.5 to 300 μg/h, about 0.75 to 200 μg/h about 1 to 100 μg/h, about 1 to100 μg/h about 1.25 to 30 μg/h, or about 0.5 to 50 μg/h and, in certainaspects about 0.5 to 200 μg/h, about 0.05 to 100 μg/h, about 0.25 to 150μg/h, about 0.5 to 50 μg/h, about 1 to 100 μg/h, about 0.75 to 200 μg/h,about 0.5 to 30 μg/h, or about 0.1 to 75 μg/h.

Additionally, in some embodiments, the nebulisers may deliver CSF-1 toinfants at a dose ranging from 0.01-1000 mg/hour of CSF-1. CSF-1 to theinfants at a dose ranging from 0.01-1000 mg/hour of CSF-1 continuouslyover 8 hours for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days or 7consecutive days. Typically, such an amount may be, in the case ofadministration to the premature infant, in the range of about 0.1 to 500mg/h, about 0.05 to 500 mg/h, about 0.2 to 400 mg/h, 0.1 to 1000 mg/h,about 0.05 to 1000 mg/h, about 0.1 to 1000 mg/h, about 0.2 to 400 mg/h,about 0.5 to 300 mg/h, about 0.75 to 200 mg/h about 1 to 100 mg/h, about1 to 100 mg/h about 1.25 to 30 mg/h, or about 0.5 to 50 mg/h and, incertain aspects about 0.5 to 200 mg/h, about 0.05 to 100 mg/h, about0.25 to 150 mg/h, about 0.5 to 50 mg/h, about 1 to 100 mg/h, about 0.75to 200 mg/h, about 0.5 to 30 mg/h, or about 0.1 to 75 mg/h.

Alternatively, in some embodiments, CSF-1 and/or a precursor, variant,analogue, derivative thereof or combination thereof, may be administeredto a premature infant directly into the bloodstream by intramuscular(i.m.), intravenous (i.v.), subcutaneous (s.c.) administration and/orcombinations thereof. The most familiar type of vascular access is aperipheral intravenous line (PIV) attached to an i.v. pump. In newborns,PIVs often may be placed in veins of the hand, foot, or scalp that mayenable delivery of CSF-1 in combination with fluids, nutrients or otherpharmaceutical agents. The PIV may enable the continuous infusion orpulse infusion of CSF-1 for hours to days. CSF-1 may be delivered by i.vinfusion to the infants at a dose ranging from 0.01-1000 μg/hour ofCSF-1 continuously over 8 hours for 1 day, 2 days, 3 days, 4 days, 5days, 6 days or 7 consecutive days. Typically, such an amount may be, inthe case of administration to the premature infant, in the range ofabout 0.1 to 500 μg/h, about 0.05 to 500 μg/h, about 0.2 to 400 μg/h,0.1 to 1000 μg/h, about 0.05 to 1000 μg/h, about 0.1 to 1000 μg/h, about0.5 to 300 μg/h, about 0.75 to 200 μg/h about 1 to 100 μg/h, about 1 to100 μg/h about 1.25 to 30 μg/h, or about 0.5 to 50 μg/h and, in certainaspects about 0.5 to 200 μg/h, about 0.05 to 100 μg/h, about 0.25 to 150μg/h, about 0.5 to 50 μg/h, about 1 to 100 μg/h, about 0.75 to 200 μg/h,about 0.5 to 30 μg/h, or about 0.1 to 75 g/h.

CSF-1 may be delivered to infants by i.v infusion at a dose ranging from0.01-1000 mg/hour of CSF-1 at a dose ranging from 0.01-1000 mg/hour ofCSF-1 continuously over 8 hours for 1 day, 2 days, 3 days, 4 days, 5days, 6 days or 7 consecutive days. CSF-1 may be delivered in the rangeof about 0.1 to 500 mg/h, about 0.05 to 500 mg/h, about 0.2 to 400 mg/h,0.1 to 1000 mg/h, about 0.05 to 1000 mg/h, about 0.1 to 1000 mg/h, about0.2 to 400 mg/h, about 0.5 to 300 mg/h, about 0.75 to 200 mg/h about 1to 100 mg/h, about 1 to 100 mg/h about 1.25 to 30 mg/h, or about 0.5 to50 mg/h and, in certain aspects about 0.5 to 200 mg/h, about 0.05 to 100mg/h, about 0.25 to 150 mg/h, about 0.5 to 50 mg/h, about 1 to 100 mg/h,about 0.75 to 200 mg/h, about 0.5 to 30 mg/h, or about 0.1 to 75 mg/h.

In some embodiments, the CSF-1, at least one precursor, at least onevariant, at least one analogue, at least one derivative, or combinationsthereof, may be administered in the form of a composition comprising acarrier (e.g. a pharmaceutically acceptable vehicle or diluent such assaline). Infants that are intubated may receive CSF-1 as an aerosol withnebulizer treatment before or in combination with another pharmaceuticalagent including but not limited to surfactants such as artificial ornatural surfactants, such as EXOSURF, PUMACTANT, KL-4, VENTICUTE.ALVEOFACT, CUROSURF, INFASURF or SURVANTA, anti-inflammatory agents orcorticosteroids, fluids for hydration, heparin, albuterol, antibiotics,ibruprofen, nutritional supplements, vitamin supplements, mineralsupplements, sildenafil, other colony stimulating factors such as G-CSFor GM-CSF, and/or IGF-I, IGF-II, HGF, EGF, or mixtures thereof.

In some embodiments, CSF-1 may be delivered to the foetus via thepregnant woman by i.v bolus injection typically at a concentration rangeof 0.1-1 g/kg body weight. The exact amount may vary depending upon avariety of factors including the relative activity, metabolic stabilityand length of action of the CSF-1, precursor, variant, analogue orderivative thereof, the route and time of administration, the degree, orlikely degree, of organ underdevelopment, and, in the case of thefoetus, the general health of the pregnant mother.

Alternatively, in some embodiments, CSF-1 may be delivered by aerosolnebulisation with or without ventilation. CSF-1 may be administered inthe nebulizer as a bolus dose before the initiation of positive pressureventilation. Alternatively, CSF-1 may be delivered by continuous feedthrough an infusion set into a nebulizer. The doses and timing for CSF-1delivery may be similar to infants described above.

In some embodiments, in pregnant warm blooded animals, such as mares,pigs, cattles, dogs and other livestock, CSF-1 may be delivered to thefoetus by bolus injection into the bloodstream of the mother usingintramuscular, intravenous, or subcutaneous administration orcombinations of the above at a concentration range typically from 0.1-1g/kg of body weight. Alternatively, in other embodiments, CSF-1 may bedelivered to such animals or to infant animals by aerosol nebulisationwith or without ventilation by administration of a bolus in thenebuliser before initiation of positive pressure or by using acontinuous feed through an infusion set into a nebuliser. The doses andtiming for CSF-1 delivery may be similar to human infants describedabove.

In some embodiments, the CSF-1, at least one precursor, at least onevariant, at least one analogue, at least one derivative, or combinationsthereof, may be administered in the form of a composition comprising acarrier (e.g. a pharmaceutically acceptable excipient, vehicle ordiluent). Such compositions may further comprise other therapeuticagents (e.g. IGF-I, IGF-II, HGF, EGF, or mixtures thereof) and may beformulated by, for example, employing conventional solid or liquidexcipients, vehicles, diluents or combinations thereof, as well aspharmaceutical additives of a type appropriate to the mode of desiredadministration (for example, excipients, binders, preservatives,stabilisers, flavours, colorants, buffers etc.). Non-limiting examplesof suitable excipients, vehicles and diluents may be found in Gennaro,Alfonso, Remington's Pharmaceutical Sciences, 18^(th) edition, MackPublishing Co. (1990), in Gennaro, Alfonso, Remington: The Science andPractice of Pharmacy, 19^(th) edition (1995) and, 20^(th) edition (2003)and/or in University of the Sciences in Philadelphia (editor) Remington:The Science and Practice of Pharmacy and 21^(st) edition (2005), theentire contents of each of which is hereby incorporated by reference.Examples of some excipients include sterile liquids, such as water andoils, including those of petroleum, animal, vegetable or syntheticorigin, such as peanut oil, soybean oil, mineral oil, sesame oil, salinesolutions, aqueous dextrose solutions, aqueous glycerol solutions,buffers, proteins such as serum albumin, amino acids such as asparticacid, glutamic acid, lysine, arginine, glycine, histidine, peptides,carbohydrates such as saccharides , polymeric additives, antimicrobialagents, sweeteners, antioxidants, antistatic agents, surfactants (e.g.,polysorbates such as “TWEEN 20” and “TWEEN 80”), lipids (e.g.,phospholipids, fatty acids), steroids (e.g., cholesterol), and chelatingagents, or combinations thereof. Examples of some buffers that may beused include salts prepared from an inorganic acid such as mineral acidsalts, such as hydrochlorides, bromides, and sulfates and salts preparedfrom an organic acid or base, such as salts of citric acid, propionicacid, malonic acid, ascorbic acid, gluconic acid, carbonic acid,tartaric acid, succinic acid, acetic acid, or phthalic acid or Tris,tromethamine hydrochloride, phosphate buffers, or combinations thereof.

The CSF-1, at least one precursor, at least one variant, at least oneanalogue, at least one derivative, or combinations thereof, may beadministered in any amount that is effective in treating complicationsarising from low birth weight in mammals, such as humans, pigs, horses,dogs or other livestock, such as in premature infants or foetuses, inpromoting organ development in the premature infant or foetus and/or inpromoting bone growth and/or enhancing bone development and/ormaturation in mammals, such as humans, pigs, horses, dogs or otherlivestock, such as in premature infants or foetuses. Typically, such anamount will be, in the case of administration to the premature infant,in the range of about 0.1 to 500 μg/kg/day, about 0.05 to 500 μg/kg/day,about 0.1 to 500 μg/kg/day, about 0.2 to 400 μg/kg/day, about 0.5 to 300μg/kg/day, about 0.75 to 200 μg/kg/day about 1 to 100 μg/kg/day, about 1to 100 μg/kg/day about 1.25 to 30 μg/kg/day, or about 0.5 to 50μg/kg/day and, in certain aspects more preferably, about 0.5 to 200μg/kg/day, about 0.05 to 100 μg/kg/day, about 0.25 to 150 μg/kg/day,about 0.5 to 50 μg/kg/day, about 1 to 100 μg/kg/day, about 0.75 to 200μg/kg/day, about 0.5 to 30 μg/kg/day, or about 0.1 to 75 μg/kg/day andfor the foetus (where administration is via the pregnant mother).However, the exact amount may substantially vary depending upon avariety of factors including, but not limited to, the relative activity,metabolic stability and length of action of the CSF-1, at least oneprecursor, at least one variant, at least one analogue, at least onederivative, or combinations thereof, the route and time ofadministration, the degree, or likely degree, of organ underdevelopment,the type and severity of complications arising from the low birthweight, the age of the foetus or premature infant and, in the case ofthe foetus, the general health of the pregnant mother.

Administration of the CSF-1, at least one precursor, at least onevariant, at least one analogue, at least one derivative, or combinationsthereof, to the premature infant may commence upon birth and continueuntil a desired level of organ development is observed. Thus, for thelung, a desired level of lung development may be achieved when thesaccular phase has been completed and/or the infant no longer requires amechanical ventilator. For the kidney, a desired level or organdevelopment may be achieved when renal function has been improved and/orthe numbers of nephrons has been increased since birth. For the foetus,administration of the CSF-1, at least one precursor, at least onevariant, at least one analogue, at least one derivative thereof, orcombinations thereof, may commence from about gestational week 4, aboutgestational week 5, about gestational week 7, about gestational week 10,about gestational week 12, about gestational week 14, or aboutgestational week 19, but in certain aspects preferably, commences afterabout week 20, about week 22, about week 24, or about week 26.

Certain embodiments disclosed also encompasses the administration of atleast one nucleic acid molecule encoding CSF-1, at least one precursor,at least one variant, at least one analogue, at least one derivativethereof, or combinations thereof, such that the CSF-1, at least oneprecursor, at least one variant, at least one analogue, at least onederivative thereof, or combinations thereof, are expressed from thenucleic acid molecule by the premature infant or foetus (and/or pregnantmother).

In certain embodiments, suitable nucleic acid molecules may be single ordouble stranded, such as mRNA, ssRNA, dsRNA, ssDNA and dsDNA. However,in certain preferred aspects, the nucleic acid molecule will be dsDNA.

The nucleic acid molecule may be incorporated into an expressionconstruct or vector in accordance with any effective method, some ofwhich are known. Typically, in certain aspects the nucleic acid moleculewill be introduced into such an expression construct or vector such thattranscription of the nucleic acid molecule is driven by a promotersequence provided by the expression construct or vector. In certainaspects it is preferred that the expression construct or vector isadapted for expression in mammalian cells, tissues or organs such aslung and/or kidney cells.

In certain aspects it is preferred that the nucleic acid molecule isincorporated into at least one viral vector such as an adenovirus,lentivirus or poxvirus vector.

The nucleic acid molecule may be administered to the premature infant orfoetus (via the pregnant mother) by any effective method, some of whichare well know (e.g. liposome-mediated transfection, or for viralvectors, viral transformation).

Administration of the at least one nucleic acid molecule encoding theCSF-1, at least one precursor, at least one variant, at least oneanalogue, at least one derivative thereof, or combinations thereof, tothe premature infant may occur upon birth, whereas for the foetus,administration of the at least one nucleic acid molecule encoding theCSF-1, at least one precursor, at least one variant, at least oneanalogue, at least one derivative thereof, or combinations thereof, mayoccur from about gestational week 4, about gestational week 5, aboutgestational week 7, about gestational week 10, about gestational week12, about gestational week 14, or about gestational week 19, but incertain aspects preferably, commences after about week 20, about week22, about week 24, or about week 26.

Certain methods disclosed are particularly suitable for increasinggrowth and/or enhancing lung maturation and nephrogenesis in the kidney.

In certain aspects disclosed, the methods of promoting growth and/orenhanced lung development and/or maturation in a premature infant orfoetus, comprise the step of administering to the infant or foetus;

at least one colony stimulating factor-1 protein (CSF-1), or at leastone precursor, at least one variant, at least one analogue, at least onederivative thereof, or combinations thereof, or

at least one nucleic acid molecule encoding said colony stimulatingfactor-1 protein (CSF-1), at least one precursor, at least one variant,at least one analogue, at least one derivative thereof, or combinationsthereof.

In certain aspects disclosed, the methods of promoting growth and/orenhanced lung development and/or maturation in a premature infant orfoetus, comprise the step of administering to the infant or foetus;

colony stimulating factor-1 protein (CSF-1), or precursor, variant,analogue, or derivative thereof or

nucleic acid molecule encoding said colony stimulating factor-1 protein(CSF-1), precursor, variant, analogue, or derivative thereof.

And, in certain aspects disclosed a method of promoting growth,maturation and/or enhanced kidney development in a premature infant orfoetus is provided, said method comprising the step of administering tothe infant or foetus;

colony stimulating factor-1 protein (CSF-1), or a precursor, variant,analogue or derivative thereof, or

a nucleic acid molecule encoding said colony stimulating factor-1protein (CSF-1), or a precursor, variant, analogue or derivativethereof.

In certain aspects disclosed, methods are provided for promoting growth,maturation and/or enhanced kidney development in a premature infant orfoetus, comprising the step of administering to the infant or foetus;

at least one colony stimulating factor-1 protein (CSF-1), at least oneprecursor, at least one variant, at least one analogue, at least onederivative thereof or combinations thereof, or

at least one nucleic acid molecule encoding said colony stimulatingfactor-1 protein (CSF-1), at least one precursor, at least one variant,at least one analogue, at least one derivative thereof, or combinationsthereof.

It is anticipated that the certain methods disclosed may be equallyapplicable to newborn non-human animals and non-human foetuses. Inparticular, it is anticipated that the methods of the invention might beused in relation to warm blooded animal, for example. But not limitedto, thoroughbred horses, stud animals and companion animals such as dogsand cats.

In order that the nature of the present inventions may be more clearlyunderstood, preferred forms thereof will now be described with referenceto the following non-limiting examples.

In some embodiments, the methods of treating complications arising fromlow birth weight in mammals, such as humans, pigs, horses, dogs or otherlivestock, such as in premature infants or foetuses, in promoting organdevelopment in the premature infant or foetus and/or in promoting bonegrowth and/or enhancing bone development and/or maturation in mammals,such as humans, pigs, horses, dogs or other livestock, such as inpremature infants or foetuses may include co-treatment with othertherapeutic modalities.

For example, in some embodiments, prior to birth the methods may includetreatment of the mother by administering to the mother

at least one colony stimulating factor-1 protein (CSF-1), at least oneprecursor, at least one variant, at least one analogue, at least onederivative thereof or combinations thereof, or

at least one nucleic acid molecule encoding said colony stimulatingfactor-1 protein (CSF-1), at least one precursor, at least one variant,at least one analogue, at least one derivative thereof, or combinationsthereof,

in combination with treatment with one or more drugs or other substancesto treat an underlying cause of low birth weight or complicationsresulting from low birth weight, such as drugs for hypertension,infections or diabetes, coticosteroids, tocolytics, nutritionalsupplements, vitamin supplements, mineral supplements, albuterol,antibiotics, heparin, other colony stimulating factors such as G-CSF orGM-CSF, surfactants such as artificial or natural surfactants, such asEXOSURF, PUMACTANT, KL-4, VENTICUTE. ALVEOFACT, CUROSURF, INFASURF orSURVANTA, IGF-I, IGF-II, HGF, EGF, sildenafil, ibuprofen or incombination with surgical treatment of the mother and/or fetus.

In some embodiments, prior to birth the treatment of the mother mayinclude treatment in a dosage form that enhances transplacental drugdelivery, such as using a liposomal form of the administered treatment.Such liposomal forms may be created having a variety of sizes, chargesand lipid compositions. In some embodiments, such liposomal forms may beanionic small unilamellar liposomes.

In some embodiments after birth, the methods herein may includeadministering to the premature or low birth weight infant

at least one colony stimulating factor-1 protein (CSF-1), at least oneprecursor, at least one variant, at least one analogue, at least onederivative thereof or combinations thereof, or

at least one nucleic acid molecule encoding said colony stimulatingfactor-1 protein (CSF-1), at least one precursor, at least one variant,at least one analogue, at least one derivative thereof, or combinationsthereof;

in combination with treatment with one or more drugs or other substancesto treat an underlying cause or complication of low birth weight, suchas co-administration of surfactant therapy, such as administration ofsurfactant, such as artificial or natural surfactants, such as EXOSURF,PUMACTANT, KL-4, VENTICUTE. ALVEOFACT, CUROSURF, INFASURF or SURVANTA,albuterol, heparin, sildenafil, ibuprofen, nutritional supplementation,vitamin and/or mineral supplementation, surgery, IGF-I, IGF-II, HGF,EGF, antibiotic therapy, other colony stimulating factors such as G-CSFor GM-CSF, other drug therapy to treat any complications of low birthweight.

In some embodiments, the CSF-1 may be provided as a kit, such as atleast one colony stimulating factor-1 protein (CSF-1), at least oneprecursor, at least one variant, at least one analogue, at least onederivative thereof or combinations thereof, or

at least one nucleic acid molecule encoding said colony stimulatingfactor-1 protein (CSF-1), at least one precursor, at least one variant,at least one analogue, at least one derivative thereof, or combinationsthereof in conjunction with instructions for administration, includingdosing instructions, adverse event information and adverse interactioninformation, other dosing equipment or coadministration therapy-relateddrugs, equipment and/or instructions.

For example, in some embodiments of the kits, at least one colonystimulating factor-1 protein (CSF-1), at least one precursor, at leastone variant, at least one analogue, at least one derivative thereof orcombinations thereof, may be provided as an ampule that may be added toa nebulisation unit or as a pre-dosed nebulisation system that attachesto a ventilator system for premature babies. In some embodiments of thekits, at least one colony stimulating factor-1 protein (CSF-1), at leastone precursor, at least one variant, at least one analogue, at least onederivative thereof or combinations thereof, may be provided in pre-dosedsyringes, with infant-specific i.v. lines and infant-sized needles orother administration devices. In some embodiments, CSF-1 kits may beprovided as a mixture of at least one colony stimulating factor-1protein (CSF-1), at least one precursor, at least one variant, at leastone analogue, at least one derivative thereof or combinations thereofwith saline, buffered saline or other diluent and/or as a mixture withother colony stimulating factors, such as G-CSF and/or CM-CSF, drugssuch as albuterol, antibiotics, or IGF-I, IGF-II, HGF, EGF, or asurfactant or mixtures thereof.

EXAMPLES Example 1 Materials and Methods Newborn Mouse Analysis

C57/B16 mice were time-mated and the mouse pups given threeintraperitoneal (i.p.) injections of recombinant human CSF-1 (ChironCorporation, Emeryville, Calif., USA) at days 1, 2 and 3 after birth.The CSF-1 was administered at a dose of 1 μg/g body weight at aconcentration of 1 μg/ml where the final volume did not exceed 50 μl perinjection. Litter mate aged-matched control mice received vehicle(phosphate buffered saline) control injections of the same volume. TheCSF-1-treated mouse pups were toe and tail clipped for identificationand returned to their mothers. The CSF-1-treated mouse pups and thecontrol-treated pups were killed at day 29 for light microscopy ofkidney and lung histology and estimation of glomerular number.

Histology

Kidney and lung tissue was taken from CSF-1-treated and litter matecontrol-treated mice, immersion fixed in 4% paraformaldehyde andprocessed on short cycle before embedding into paraffin for histologicalanalysis.

The paraffin-embedded kidneys were each sectioned at 4 microns using amicrotome (Leitz Wetzlar, Germany), and the sections were then placed onpoly-L-lysine slides and left to adhere for 3 hours at 70° C. The slideswere dewaxed in xylene, rehydrated through graded alcohols to waterbefore staining with haematoxylin and eosin by standard methods.

Stereological Assessment of Glomerular Number and Kidney Volume

At day 29, mice with/without delivery of CSF-1 were killed and theirkidneys removed and processed for methacrylate embedding and subsequentstereological counting. The processing involved placing the kidneys into10% formalin for 48 hours, 70% ethanol overnight, and then three onehour washes with 100% ethanol followed by butanol overnight and 72 hoursin infiltration solution (Technovik 7100, Electron Microscopy Sciences,QLD). Kidneys were then embedded in methacrylate resin (Technovik 7100)and left to set for three days. Once set, the backs of blocks were madeusing Technovik 3040 solution (Technovik, Electron Microscopy Sciences,QLD) and allowed to set for one hour. Sections were then serialsectioned at 20 μm using a microtome, and every 10th and 11th sectionwas collected beginning at a random number.

Sections were stained using periodic acid-Schiff (PAS) staining, howeverthe time in reagents was extended compared to a standard paraffinprotocol, due to the slow rate of penetration through the resin.

Stereological counting was performed firstly on a microfische reader todetermine kidney volume using the Cavalieri Principal (Kett, M M et al.,1996). Complete sections were used for nephron number estimation using aphysical dissector/fractionator combination (Bertram, J F, 1995)—wherebypairs of sections are projected onto an unbiased 2×2 cm counting grid.Grid points that lay upon kidney tissue, glomeruli and renal corpuscleswere tabulated, as were glomeruli disappearing events across each slide.Using a formula, the nephron number was estimated.

Results and Discussion

The administration of CSF-1 (1 μg/g body weight) was given to mouse pupsat Day 1, 2 and 3 after birth. In CSF-1-treated mice, there was anoverall increase in body weight (FIG. 1) and individual kidney weights(FIG. 2). Mice killed at day 29 we observed to have a 37% increase inoverall body weight compared to age-matched litter mates. CSF-1-treatedmice had a 27% and 33% increase in left and right kidney weights,respectively, at the same timepoint. The mice with CSF-1 treatment werealso found to have a 29% increase in the number of kidney glomerulicompared to age-matched litter mate control mice treated with phosphatebuffered saline (FIG. 3). This demonstrates that CSF-1 can promoteincreased nephrogenesis that is associated with increased kidney growth.

FIG. 4 demonstrates the histology of the kidneys and lungs from CSF-1(FIG. 4B, D, F) and control-treated (FIG. 4A, C, E) mice at day 29.There were no obvious structural abnormalities observed in the kidneysfrom mice following CSF-1 injection compared to control animals. On theother hand, the lungs from CSF-1-treated mice appeared more developed;in particular, the alveolar wall appeared to be thinner and lesscellular, and there was less connective tissue compared to litter matecontrol animals. This corresponds to a greater degree of alveolarisationin the CSF-1-treated animals.

In conclusion, CSF-1 was observed to have a growth promoting effect ontotal and individual kidney weights when administered systemically tonewborn mice. This increase in kidney weight in the CSF-1 treated micewas associated with increased nephrogenesis. In addition, the lungs ofthe CSF-1 treated mice appeared more differentiated compared to controllitter mate animals. Therefore, it is considered that CSF-1 showsconsiderable promise for the treatment of pregnant mothers at risk ofpremature delivery, as well as in the treatment of premature babies withthe objective to promote growth and maturation of the lungs and kidneysto prevent associated complications and disorders.

Throughout this specification the word “comprise”, or variations such as“comprises” or “comprising”, will be understood to imply the inclusionof a stated element, integer or step, or group of elements, integers orsteps, but not the exclusion of any other element, integer or step, orgroup of elements, integers or steps.

All publications mentioned in this specification are herein incorporatedby reference. Any discussion of documents, acts, materials, devices,articles or the like which has been included in the presentspecification is solely for the purpose of providing a context to thedisclosed embodiments. It is not to be taken as an admission that any orall of these matters form part of the prior art base or were commongeneral knowledge in the field relevant to the present inventions as itexisted in Australia or elsewhere before the priority date of each claimof this application.

It will be appreciated by persons skilled in the art that numerousvariations and/or modifications may be made to the disclosed embodimentsas shown herein without departing from the spirit or scope of theinventions as disclosed. The present embodiments are, therefore, to beconsidered in all respects as illustrative and not restrictive.

1. A composition for the treatment of complications arising from or related to low birth weight in a mammal comprising: a low birth weight complications-reducing or-limiting amount of at least one colony stimulating factor-1 protein (CSF-1), or at least one precursor, variant, analogue or derivative thereof, or a therapeutically effective amount of at least one nucleic acid molecule encoding a low birth weight complications-reducing or-limiting amount of said at least one colony stimulating factor-1 protein (CSF-1), or at least one precursor, variant, analogue or derivative thereof; and a pharmaceutically acceptable excipient, vehicle or diluent.
 2. A composition for promoting organ development in a mammal comprising a mammal organ development-enhancing amount of colony stimulating factor-1 protein (CSF-1), or a precursor, variant, analogue or derivative thereof, or a therapeutically effective amount of a nucleic acid molecule encoding a premature infant or foetus organ development-enhancing amount of said colony stimulating factor-1 protein (CSF-1), or a precursor, variant, analogue or derivative thereof; and a pharmaceutically acceptable excipient, vehicle or diluent.
 3. A composition for promoting lung growth and/or enhancing lung development and/or maturation in a mammal comprising a mammal lung growth promoting and/or lung development and/or maturation-enhancing amount of colony stimulating factor-1 protein (CSF-1), or a precursor, variant, analogue or derivative therefore, or a therapeutically effective amount of a nucleic acid molecule encoding a mammal lung growth promoting and/or lung development and/or maturation-enhancing amount of said colony stimulating factor-1 protein (CSF-1), or a precursor, variant, analogue or derivative thereof; and a pharmaceutically acceptable excipient, vehicle or diluent.
 4. A composition for promoting growth, maturation and/or enhancing kidney development in a mammal comprising a mammal kidney growth maturation and/or development-enhancing amount of at least one colony stimulating factor-1 protein (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof, or a therapeutically effective amount of at least one nucleic acid molecule encoding a mammal kidney growth maturation and/or development-enhancing amount of at least one said colony stimulating factor-1 protein (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof; and a pharmaceutically acceptable excipient, vehicle or diluent.
 5. A composition for promoting bone growth and/or enhancing bone development and/or maturation in a mammal comprising a mammal bone growth promoting and/or bone development and/or maturation-enhancing amount of at least one colony stimulating factor-1 protein (CSF-1), and/or a precursor, variant, analogue, derivative thereof or combination thereof, or a therapeutically effective amount of at least one nucleic acid molecule encoding a mammal bone growth promoting and/or bone development and/or maturation-enhancing amount of said at least one colony stimulating factor-1 protein (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof; and a pharmaceutically acceptable excipient, vehicle or diluent.
 6. The composition according to claim 1, wherein said mammal comprises a premature infant, a low birth weight infant or a foetus.
 7. A method of treating complications arising from or related to low birth weight in mammals comprising administering to said mammals: at least one colony stimulating factor-1 protein (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof, or at least one nucleic acid molecule encoding said at least one colony stimulating factor-1 protein (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof.
 8. A method of promoting organ development and/or maturation in mammals comprising administering to the mammal; colony stimulating factor-1 protein (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof or a nucleic acid molecule encoding said colony stimulating factor-1 protein (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof.
 9. A method of promoting lung growth and/or enhancing lung development and/or maturation in mammals, comprising administering to said mammals: colony stimulating factor-1 protein (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof, or a nucleic acid molecule encoding said colony stimulating factor-1 protein (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof.
 10. A method of promoting growth, maturation and/or enhancing kidney development mammals comprising administering to said mammals: colony stimulating factor-1 protein(CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof, or a nucleic acid molecule encoding said colony stimulating factor-1 protein (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof.
 11. A method of promoting bone growth and/or enhancing bone development and/or maturation in mammals comprising administering to said mammals: colony stimulating factor-1 protein (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof or a nucleic acid molecule encoding said colony stimulating factor-1 (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination.
 12. The method of according to claim 7 wherein said mammal comprises a premature infant, a low birth weight infant or a foetus.
 13. A method of promoting lung growth and/or enhancing lung development and/or maturation in mammals, comprising administering to said mammals: colony stimulating factor-1 protein (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof, or a nucleic acid molecule encoding said colony stimulating factor-1 protein (CSF-1), and/or at least one precursor, variant, analogue, derivative thereof or combination thereof; and at least one surfactant.
 14. A composition for promoting lung growth and/or enhancing lung development and/or maturation in a premature infant, in a low birth weight infant or in a foetus comprising: a premature infant, low birth weight infant or foetus lung growth promoting and/or lung development and/or maturation-enhancing amount of colony stimulating factor-1 protein (CSF-1); and a pharmaceutically acceptable excipient, vehicle or diluent.
 15. A composition for promoting growth, maturation and/or enhancing kidney development in a premature infant, in a low birth weight infant or in a foetus comprising: a premature infant, low birth weight infant or foetus kidney growth maturation and/or development-enhancing amount of colony stimulating factor-1 protein (CSF-1); and a pharmaceutically acceptable excipient, vehicle or diluent.
 16. A method of promoting lung growth and/or enhancing lung development and/or maturation in a premature infant, in a low birth weight infant or in a foetus, comprising administering to said premature infant, low birth weight infant or foetus: colony stimulating factor-1 protein (CSF-1).
 17. A method of promoting growth, maturation and/or enhancing kidney development in a premature infant, in a low birth weight infant or in a foetus comprising administering to said premature infant, low birth weight infant or foetus: colony stimulating factor-1 protein (CSF-1). 